Ecology and Evolution

Understanding how closely related species coexist in highly seasonal and unpredictable environments is central to studies of ecological differentiation and niche partitioning. We investigated the feeding ecology of sympatric populations of Microcebus murinus and M. griseorufus within a contact zone in Andohahela National Park, southeastern Madagascar, across dry and wet seasons. Using a combination of direct behavioral observations (1,611 feeding records), fecal sample analyses (n = 56), and vegetation phenology surveys, we quantified dietary composition, seasonal shifts in resource use, and habitat-related variation. Seasonal changes in diet were pronounced, with dry-season feeding dominated by exudates and wet-season diets incorporating greater proportions of fruit and flowers, closely tracking phenological patterns at both sites. Diets of both species were dominated by plant resources, but consistent interspecific differences in dietary strategy were evident. Although both species consumed comparable proportions of insect prey, M. murinus showed pronounced wet-season increases in the use of high-sugar, carbohydrate-rich floral resources (15.9%) and hemipteran-associated honeydew (30.6%). In contrast, M. griseorufus relied more consistently on predictable exudates throughout the year. Fecal analyses supported observational data but revealed differences in the detectability of dietary components, with increased representation of invertebrates in the wet season and seeds in the dry season. These results indicate substantial dietary overlap but consistent differences in resource use, suggesting that coexistence is facilitated by fine-scale trophic differentiation within a broadly shared omnivorous niche. Such subtle but persistent differences in feeding strategy likely reduce competitive overlap and enable continued sympatry in a climatically variable and resource-limited system.

Population dynamics and their component vital rates may be driven by weather, climate teleconnections between sea and air (e.g. ENSO), or biotic interactions. These drivers operate directly or indirectly and on different temporal scales. We used a Bayesian structural equation model to characterize effects among weather, climate, and incidental intraguild predation (IGP) on the butterfly Euphydryas gillettii's vital rate of pre-diapause survival, using an 18 year dataset. IGP was a major determinant of pre-diapause survival, along with direct and indirect effects of weather and spring climate teleconnections. The direction of climate effects was reversed when mediated through IGP. Our analysis illustrates the need for sequential hypotheses to capture the cascading effects of abiotic factors via biotic interactions. Using sequential hypotheses addresses the debate on weather -- climate teleconnection roles by disentangling their contributions from one another. Finally, vital rates must be decomposed to component rates in order to detect their drivers.

Heritable bacterial symbionts are pervasive in terrestrial arthropods, often imposing reproductive manipulations to promote their own spread within host populations. Co-infections are common, potentially allowing symbiont co-infectors to hitchhike through a host population. However, adverse thermal conditions can disrupt these communities, particularly when co-infectors vary in their thermal sensitivity. We used a multi-generation experiment to test whether warm (29 {degrees}C) conditions disrupted spread of heritable symbionts through uninfected populations of the spider, Mermessus fradeorum. We tested two common infection combinations: a single infection with a cytoplasmic incompatibility (CI) inducing Rickettsiella or a feminizing co-infection that included a feminizing Wolbachia, the same Rickettsiella, and up to three apparent hitchhikers (two additional Wolbachia strains and Tisiphia). We initiated replicate populations with 1/3 of one infection type and 2/3 uninfected spiders, evaluating population infection rate over 5 spider generations under different temperature regimes. Under cool (21{degrees}C) conditions, Wolbachia feminization drove co-infection to 88% and Rickettsiella CI drove single infection to 83% of host populations. Vertical transmission for all symbionts was high (97-99%) and hitchhiking symbionts also spread effectively. Under warm conditions, feminization and CI efficacy were reduced, and symbionts suffered variably reduced vertical transmission. Warm conditions ultimately destroyed the co-infecting symbiont consortium and impeded symbiont spread. On its own, though, Rickettsiella was still able to increase, despite reduced strength of CI. We hypothesize that contrasting tensions between feminizing spread of the symbiont consortium versus environmentally driven loss of function and transmission may explain observed patterns of mixed infections in field populations of this spider.

Animals reproduce across a spectrum from oviparity to viviparity. Internal gestation and live birth have inherent advantages despite their costs. However, comparative analyses of key drivers, such as resilience to dehydration, lack taxonomic breadth. In this study, we assessed whether live birth improves the ability to proliferate in environments with limited access to water. To do so, we used the viviparous cockroach, Diploptera punctata, as a model of viviparity to assess dehydration-induced damage during extended periods of water deprivation. During pregnancy, maternal survival did not decline during dry periods compared with non-pregnant individuals, suggesting that pregnancy has minimal impact on resistance to dehydration stress. When mothers are deprived of water for 2 weeks, they show an increase in osmolality after one week, while their embryos show no change in osmolality until after two weeks, at which point abortions occur. Periods of dehydration increased the duration of pregnancy, likely due to a slower production of in utero milk, but there was no change in the size of progeny. After birth, viviparous D. punctata newborns showed increased survival under dry conditions compared to two other ovoviviparous species, which are much smaller and dehydrate more quickly. The combined impact of increased dehydration resistance during pregnancy and during the first independent phase indicates a benefit of prolonged viviparity in Diploptera punctata. This study provides evidence that viviparity in animal systems could enable progeny survival during drought.

Phenotypic plasticity, the ability of a single genotype to produce alternative phenotypes in response to environmental cues, is a key driver of evolutionary change. In the water strider Microvelia longipes, males display remarkable continuous variation in hindleg length, a sexually selected trait used as a weapon in male/male contests for access to females. To determine whether DNA methylation mediates this environmentally induced phenotypic variation, we used three inbred lines of M. longipes that differ in mean hindleg length, body size, and allometric coefficients. We performed whole-genome bisulfite sequencing on adult males and females from all lines, and tested the effect of nutritional treatment on DNA methylation patterns. Our analysis identified 12,684,876 CpG 12% of which were methylated. This global level of DNA methylation is among the highest reported in insects. DNA methylation was predominantly concentrated within or near gene bodies (77% of methylated CpGs), consistent with patterns observed in other insects. Unsupervised clustering and principal component analyses revealed that methylation patterns differed significantly between genetic lines but showed minimal differences between sexes, indicating a strong genetic influence. Most surprisingly, despite nutrition having a pronounced effect on leg length, we observed no significant changes in DNA methylation in response to dietary treatment. These results show that in M. longipes, DNA methylation patterns are largely stable across environmental conditions and primarily determined by genetic background. This challenges the common assumption that DNA methylation universally mediates environmentally induced phenotypic plasticity and suggests that other epigenetic mechanisms, such as histone modifications or non-coding RNAs, may play a more direct role in regulating continuous plastic traits. Our study underscores the complexity of epigenetic regulation and highlights the need for broader investigation of molecular pathways to fully understand the molecular basis of phenotypic variation in natural populations.

Copper is an essential micronutrient in most organisms that becomes toxic in large quantities. Repeated or prolonged sub-lethal exposure can lead to evolved resistance to copper toxicity over many generations, which may result in trade-offs between energetically expensive detoxification mechanisms and fitness. Alternatively, evolved resistance to chemical stressors may lead to correlated changes in other traits. This study focuses on a population of flies for which artificial selection for copper resistance led to an increase in both copper resistance and longevity. The apparent off-target benefit of copper selection on one component of fitness led us to investigate differences in fecundity and developmental viability in copper resistant and copper sensitive, non-selected populations. We assessed the effect of copper selection and copper exposure on multiple aspects of fecundity over the lifespans of females from the non-selected and copper-selected populations. Our study corroborated previously observed increased longevity in copper-selected flies. Controlling for variation in lifespan, copper-resistant females had comparable age-matched fecundity to copper-sensitive females and benefitted from increased longevity with higher lifetime fecundity. Overall, copper exposure negatively affected egg quality, but we found no difference in this trait between the copper-resistant and sensitive populations. Further, we found developmental viability under copper stress was significantly higher for eggs laid by copper-resistant females. Overall, we determined that copper resistant flies experienced a fitness benefit through both lifespan and fecundity. Costs of maintaining copper resistance may be associated with energetic costs, but these trade-offs may not always manifest in reproductive or lifespan fitness costs.

This study aimed to examine entomologists' perceptions of equity, inclusion, and exclusion within the discipline, identifying perceived challenges and proposed solutions for advancing equity in the field. The present study surveyed 47 self-identified entomologists living in the United States in 2023. Using a mixed-methods design, the study examined entomologists' perceptions of inclusion and exclusion within the discipline through qualitative and quantitative measures. Respondents highlighted needs for race-conscious funding opportunities, comprehensive inclusion efforts through geographically diverse outreach, and access to role models and mentors with similar identities to future entomologists. Findings are discussed in relation to a smaller 2013 study on recruitment and retention of entomologists of color, which offers preliminary historical context. The 2013 respondents emphasized intrinsic and age-based barriers to recruitment (e.g., lack of interest, limited K-12 outreach), the 2023 findings point toward structural inequities and retention needs (e.g., systemic exclusion). The 2013 comparison is interpreted as exploratory given differences in sample size and scope between the two studies. The 2023 study highlights evolving perceptions of persistent inequities in entomology and identifies opportunities to build a more inclusive and representative discipline.

PurposeDiscussions surrounding the biomedical faculty job market often focus on applicant competitiveness and external metrics such as number of publications and funding records. Consequently, there is typically less discussion about applicant readiness, the point at which applicants perceive themselves as prepared to enter the market. Since 2018 our group, the Faculty Job Market Collaboration (FJMC), has conducted annual end-of-cycle surveys of biomedical faculty job applicants, producing the largest longitudinal dataset on this process to date. MethodsWe employed a mixed-methods design examining faculty applicants in biological science fields in North America. Regression analyses were conducted on a longitudinal dataset of 729 respondents across multiple hiring cycles. To determine how applicants evaluated their own preparation, qualitative interviews were conducted with a separate cohort of biomedical postdoctoral applicants during the 2024-2026 job cycles. ResultsOur findings demonstrate that rather than depending on a single quantitative threshold, readiness is a multifaceted construct shaped by actionable and interpersonal drivers. Key factors influencing an applicants perceived readiness include taking agency to submit applications, receiving explicit support from a mentor, incorporating strategic use of artificial intelligence tools into application preparation, and their career stage. ConclusionBy distinguishing individual readiness from systemic assumptions of market competitiveness, this study highlights a blind spot in academic workforce development. Our results suggest that applicants can achieve readiness and successful outcomes through different combinations of support, strategy, and timing rather than a uniform metric profile. By integrating quantitative and qualitative data, our study provides an evidence-based framework for understanding applicant readiness and offers practical guidance to help trainees navigate the increasingly competitive academic job market. Teaser TextOur mixed-model analysis of the biomedical faculty job market is designed to help prospective faculty candidates assess their readiness to enter the job market. By integrating multiple indicators of academic productivity, funding success, and professional experience, our study provides evidence-based benchmarks that can guide applicants in evaluating their competitiveness and identifying areas for further development before pursuing faculty positions.

Riparian environments are characterised by a high diversity of arthropod species, linking the aquatic with the terrestrial ecosystem. One of the dominant arthropod predators in this ecotone, carabid beetles are of particular interest as they may also act as facultative scavengers, feeding on carrion deposited along the shoreline. To test whether carabids feed on carrion, we examined the feeding preferences of the abundant riparian carabid Bembidion elongatum in a laboratory feeding trial, offering freshly killed and 24 hours post mortem Drosophila melanogaster. We subsequently assessed the detection probability of ribosomal prey RNA and DNA in predator gut contents using Drosophila-specific RT-PCR and PCR assays, and quantified nucleotide abundance by quantitative real-time PCR at 0, 3, 6, and 12 hours post-feeding. In the feeding experiment, B. elongatum showed a significant preference for fresh over carrion prey. Following consumption, the quantities of prey DNA and RNA in the predators gut declined over a 12-hour post-feeding period. However, no differences were detected in prey DNA or RNA quantities between individuals fed fresh prey and those fed carrion. Only immediately after consumption was the DNA:RNA ratio significantly lower in individuals fed fresh prey compared to those fed carrion while this difference was not observed at later time points. Overall, our results indicate that ingested ribosomal prey RNA in predators is present in high quantities, and that the DNA:RNA ratio is not a suitable indicator for distinguishing between consumption of carrion and fresh prey.

The ability to model the complex and uncertain population dynamics of endangered species has improved dramatically in recent decades. However, approaches to identify optimal decisions often require a simplified representation of population dynamics. This leads to a conundrum where managers may be unsure about the output of dynamic decision models because they rely on simplified assumptions of the underlying population dynamics. Here, by pairing integrated population models (IPM) that synthesize diverse ecological data with deep reinforcement learning (DRL) capable of optimizing decisions with high-dimensional uncertainty, we introduce a framework that delivers data-driven and ecologically detailed adaptive management strategies. We demonstrate its utility through application to the supplementation program for the endangered Rio Grande silvery minnow. Using our IPM-DRL framework, we developed an adaptive decision model that selects production and distribution decisions of the supplementation program in response to the observed demographic, hydrological, and genetic environment. The decision model outperformed all heuristic approaches in the simulation across management objectives that weighed persistence and effective population size-related genetic impact differently. For example, the currently deployed supplementation strategy performed 5.3% worse than the decision model under the persistence-focused objective scoring and 185% worse under the genetics-focused one. Analysis of the models decisions in relation to demographic and environmental covariates revealed that minimum sub-population size and total population size were primary drivers of the models decisions. The results demonstrate that the IPM-DRL framework offers a high-performing and interpretable decision-support tool for managing endangered species. SignificanceConservation problems, like imperiled species management, are often challenging because the system dynamics are complex and uncertain. We demonstrate how combining an integrated population model that infers key demographic processes from noisy ecological data with a deep reinforcement learning framework that optimizes management actions addresses these challenges by generating high-performing supplementation strategies for a conservation-dependent species. Our approach embeds two decades of monitoring data within a multi-objective decision-making environment that accounts for ecological uncertainty. The result is a generalizable framework that links ecological inference directly to actionable policy outcomes, enabling scientists and managers to move beyond describing system states and processes toward identifying optimal management actions.

Phenotypic differences among populations can arise through heritable genetic divergence, phenotypic plasticity, or both, making it difficult to determine whether trait-environment correlations observed in nature reflect adaptive evolution. Within threespine stickleback (Gasterosteus aculeatus) studies, numerous document morphological differences among allopatric-, parapatric-, and even sympatric populations. These phenotypic differences among populations are often correlated with diet and lake habitat (e.g., lake size), suggesting an adaptive value to the population differences. However, many studies of ecomorphological divergence in stickleback use wild-caught stickleback, which may differ due to evolution or plasticity. Although common garden experiments have confirmed that population differences can be heritable, such experiments typically entail small numbers of populations. Consequently, we still do not know to what extent well-known trait-environment correlations in stickleback are a result of evolution. To address this gap, we reared stickleback embryos from 27 lake populations on Vancouver Island, in a laboratory environment. Morphological differences among populations persist in common-garden fish, confirming a large role for divergent evolution. These heritable differences were associated with environmental variation among lakes, implying an adaptive value. However, some well-known trait-environment relationships in stickleback did not persist in common-garden fish and may be primarily plastic.

Predation is a major selective force shaping lemur behaviour and population dynamics, yet direct observations remain rare, particularly for small nocturnal species. Here, we document predation on mouse lemurs (Microcebus spp., Cheirogaleidae) in Andohahela National Park, south-eastern Madagascar, and using complementary evidence from radio-telemetry and owl pellet analyses. Two radio-collared individuals were confirmed as prey of endemic snakes, Ithycyphus oursi and Madagascarophis meridionalis (both Pseudoxyrhophiidae), representing the first documented records of Microcebus predation by these species. Examination of owl pellets and prey remains from three sites and from three owl species revealed a single predation event by Asio madagascariensis (Strigidae), likely involving M. tanosi, no evidence of mouse lemur predation by Tyto alba (Tytonidae) despite high local prey availability, and a single and first predation record from Athene superciliaris (Strigidae). Material from A. superciliaris roosts was otherwise dominated by invertebrates, indicating that primate predation is likely opportunistic. Together, these findings expand the known predator guild of mouse lemurs and suggest that snake predation is rarely detected, and its contribution to mouse lemur mortality and population dynamics is likely underestimated. Our findings demonstrate how combining behavioural field observations with dietary evidence can uncover otherwise undetected predation events and clarify predator prey relationships in nocturnal primates.

1. Most species are neither discovered nor named, let alone included in analyses that require biological information such as trait measurements, images, ecological information and genome-scale data. Specimen-level DNA barcoding can help discover many of these species rapidly, but everything beyond discovery requires vouchers organized into putative species. Yet, existing barcoding workflows lack efficient techniques for voucher recovery, creating a post-barcoding bottleneck that limits the ability of converting barcoded specimens into biological knowledge. 2. Here we present a low-cost, open-source workflow consisting of two stages. The first safeguards barcoded specimens by separating them from DNA extracts and transferring them from microplates into ethanol-filled glass vials. The second converts the resulting voucher collection into a searchable physical resource by linking barcode-derived molecular Operational Taxonomic Unit (mOTU) assignments to vial positions and enabling specimens to be sorted into putative species either manually or automatically using a newly developed open-access robot (SORTER). 3. We evaluated the workflow using 2,024 insect specimens distributed across 21 96-well plates. For the first stage, DNA separation and specimen transfer required approximately 15 minutes per plate. For the second stage, MOTUmapper generated retrieval coordinates in a few seconds, after which the 2,024 vouchers belonging to the 452 putative species could be recovered manually in 5 days or with SORTER in 5 hours. Throughout both stages, specimen identities remained linked to barcode sequences, metadata and storage positions. 4. Vouchers are the Rosetta stones of biology because they connect different kinds of data to the same specimens. By safeguarding these vouchers and making them searchable, the workflow converts barcode projects from one-time molecular surveys into reusable resources for ecological and evolutionary research.

High-throughput amplicon sequencing has transformed microsatellite (STR) genotyping by overcoming many of the limitations of fragment-length analysis, enabling more accurate, cost-effective, and standardized genotyping. Yet, protocols specifically designed for high-throughput sequencing (HTS)-based STR genotyping from low-template and degraded DNA remain scarce, despite the prevalence of these challenging sample types in ecological and conservation contexts. We present a methodology for the de novo development of robust STR multiplex panels together with a laboratory protocol for efficient and reliable STR genotyping by sequencing with low quantity and quality DNA samples. The protocol comprises (i) an automated bioinformatic pipeline to design large sets of short tetranucleotide markers optimized for multiplex amplicon sequencing of degraded and low-template DNA; (ii) guidelines for efficient in vitro optimization of multiplex amplification using directly low quantity/quality template DNA; and (iii) a library preparation procedure that improves detection of low-level allele signal while enabling quality assessment of STR amplicon sequencing under limiting DNA conditions. We demonstrate the approach by developing and validating STR panels for non-invasive genotyping of three large carnivore species: a 44-plex for the grey wolf (Canis lupus), a 41-plex for the Eurasian lynx (Lynx lynx), and a 30-plex for the brown bear (Ursus arctos). Multiplex performance was high, with [≥]91% of samples successfully genotyped at [≥]50% of loci (allele size range 28-110 bp across panels) and correctly assigned to known individuals, negligible levels of noise in the controls, and high discriminatory power (PIDsibs [≤]2.4 x 1e-12), also owing to sequence variation among same-length alleles at 15-50% of loci. The approach is broadly applicable to animal and plant species, a wide range of sample types, and large-scale analysis such as genetic monitoring. Our study reinforces the value of STR amplicon sequencing for ecological and conservation applications while highlighting the importance of marker design and laboratory workflows tailored to HTS-based genotyping for accurate and efficient implementation.

Wildlife diseases cause well-documented and often dramatic reductions in host survival. However, the impact of infectious diseases on host reproduction remains understudied, especially with respect to effects of prior and/or current pathogen exposure on reproductive development. Here we experimentally tested how prior and/or current infection with a common bacterial pathogen, Mycoplasma gallisepticum ( MG), alters reproductive development for female versus male house finches (Haemorhous mexicanus). Finches were inoculated with either MG or sterile media while in wintering condition and subsequently received one of these treatments while in breeding condition. In females, MG exposure had both immediate and carry-over effects on reproduction: controls had higher odds of laying eggs compared to females inoculated with MG in spring only, higher odds than females inoculated in both winter and spring, and higher odds than females given MG in the winter only. Conversely, breeding-condition males inoculated with MG in spring had higher testosterone levels than males receiving only control inoculations, and there were no carryover effects of winter MG inoculation or inoculations during both seasons on testosterone. Sex bias in the reproductive impacts of infectious diseases may have important knock-on effects on the epidemiology and population-regulating effects of pathogens, thereby warranting further study.

Machine learning models can be used to analyze large bioacoustics datasets and explore variation due to geography or habitat. We find that in the monotypic Lark Sparrow (Chondestes grammacus), both environmental variables and geographic distance influence song variation in this species. Bird song is an important method of communication within avian species. The variation in bird song within a species can be due to a variety of factors, including genetic drift and isolation by distance. However, it remains unclear in species with wide ranges how environmental factors in particular can cause changes to the song. In this study, the song C. grammacus was analyzed via machine learning to determine if it had significant variation based on multiple geographical metrics. We trained a convolutional neural network to segment individual syllables of 91 C. grammacus recordings, then extracted song characteristics. We found that ecoregion and state explain variation in C. grammacus songs. Our results demonstrate the efficacy of using machine learning models to analyze large datasets, as well as the impact that ecogeographic variation has on song variance.

Many insect populations are experiencing thermal stress as a result of global change, making it imperative to investigate how their relationship with other organisms will be impacted by heat disturbances. Microbial symbionts, such as bacteria, have the potential to enhance or inhibit an insect's thermal tolerance. Solitary bee larvae host bacteria within their food stores ("pollen provisions"), which have been shown to benefit survival and development; however, it is unclear how heatwaves brought about by climate change will impact their relationships with these bacterial partners. In this study, we subjected blue orchard bee (Osmia lignaria) eggs and larvae to a 4-day heatwave (35 {degrees}C daytime:22 {degrees}C nighttime) or kept them at control temperatures (25 {degrees}C daytime:15 {degrees}C nighttime), then returned all bees to control temperatures for a 5-day recovery period. We assessed bacterial communities within pollen provisions and larval development stage pre-heatwave (Day 0), immediately post-heatwave (Day 4), and following the recovery period (Day 9). Bacterial community composition, diversity, and abundance were resilient to heat stress, but larval bees developed faster when subjected to a heatwave. This finding refutes the hypothesis that bacteria within pollen provisions modulate blue orchard bee responses to heat, suggesting instead that developmental effects could be more largely shaped by bee physiology or interactions with microorganisms other than bacteria.

Replication is central to most experimental and sampling designs, increasing inferential power and capturing fine-scale data heterogeneity. However, its importance remains poorly evaluated in some ecological and evolutionary settings. This is the case of metabarcoding studies using DNA recovered from sedimentary archives, in which biological signals may integrate ecological information through depositional and burial processes, and are often inferred from a single sediment core per site. Here, we evaluated the effect of different types of replication using sedimentary DNA (sedaDNA) metabarcoding data from two genetic markers (mitochondrial COI and nuclear 18S), under a nested sampling design. The design included three intertidal sites, three spatially separated sediment cores per site (biological replicates), two sediment depth horizons per core, and eight PCR (technical) replicates per sediment sample. Variance partitioning showed that site identity and sediment age group together explained >70% of the variation in beta diversity, indicating that among-site spatial variation and stratigraphic variation were the dominant drivers of community composition. In contrast, variation among different cores within sites was small and non-significant (<5%). Among PCR replicates from the same sediment sample, richness varied substantially, whereas Shannon diversity was more consistent. Despite this variability, differences in community composition among technical replicates remained smaller than among biological replicates and site identity, indicating limited influence on broader ecological patterns. Community composition was highly similar among replicate cores within sites, consistent with stratigraphic coherence. These results indicate limited within-site heterogeneity and suggest that, under stratigraphically coherent conditions, increasing biological replication may yield limited additional information, whereas enhancing technical replication and stratigraphic resolution can improve ecological inference from sedaDNA metabarcoding datasets.

Flow regulation for hydropower production affects stream ecosystems through decreased connectivity and changed timing and magnitude of flows. Hydropeaking, a form of regulation in which infrequent large peak flows are replaced with frequent small peaks, increases riparian erosion and causes water and drought stress for riparian vegetation. Hydropeaking is likely to affect soil seed bank (SSB) formation and composition, while SSBs are important sources of self-restoration should a systems flow regulation be relaxed. We tested how hydropeaking intensity affects the size and composition of SSBs, including the functionally important group of large graminoids, and by calculating Ellenberg values for Moisture, Light and Soil disturbance. SSB samples were taken at 15 riparian zones across central and northern Sweden. Each site was regulated, but sites differed in their hydropeaking intensities. SSBs were subjected to a seedling emergence experiment, from which over 700 seedlings from 53 taxa emerged. We found that hydropeaking intensity affects the composition of soil seed banks on multiple levels. Seedling density was negatively correlated with hydropeaking intensity at the sites where samples were taken. SSB richness varied (two to eighteen species per site) and was not affected by hydropeaking intensity. The proportion of large graminoids in the seed bank showed a near-significant decrease with increasing hydropeaking intensity, and community-weighted means for Moisture, Light and Soil disturbance increased (non-significantly) with increasing intensity. Our results suggest that riparian SSBs, should flow regulation be relaxed or ceased, are not sufficient for self-restoration of functional riparian vegetation. Seeds of large graminoids and species that are tolerant to drought in the germination stage are less present in riparian SSBs of heavily-regulated streams. Supply of seeds of these groups, or even planting, may need to be considered when changes in flow management are implemented. HighlightsO_LIHydropeaking negatively affects riparian soil seed banks (SSBs) in Sweden C_LIO_LISSB size slightly decreases with hydropeaking intensity, but richness does not change C_LIO_LIThe proportion of large graminoid seeds in SSBs decreases with hydropeaking intensity C_LIO_LIRiparian SSBs from less-impacted sites have most potential for self-restoration C_LI

The red fox, Vulpes vulpes, is abundant in England and can exert limiting effects on their avian and mammalian prey. Large-scale gamebird releases in the UK may be sustaining high predator numbers, leading to greater predation on other prey species, especially once gamebird stocks are depleted. However, little is known how gamebird release affects the broader diet of the red fox. Here we used DNA metabarcoding to assess the diet of foxes from 18 agricultural estates in Southern England, 10 of which released large numbers of gamebirds (red-legged partridges - Alectoris rufus, and common pheasants - Phasianus colchicus) and 8 which did not. Scats were collected over one year, allowing for seasonal investigation of the foxes' diet. We investigated the vertebrate species consumed and compared the non-gamebird dietary diversity and composition between release and non-release estates and across seasons. The field vole (Microtus agrestis) was found to be the most frequently predated species overall. Brown hares and field voles were detected more on release sites, while bank voles and dog faeces were detected more on non-release sites. We found little evidence that foxes predate ground nesting birds or other species of concern. The dietary diversity was significantly lower on estates that released gamebirds, and this difference was most notable during the post-shoot, spring months (February to April). On both estate types, diversity was highest in the summer months. The altered predatory behaviour due to gamebird release is likely to affect the populations of the non-game prey of the red fox and this should be considered when designing policies regarding gamebird management and biodiversity conservation. Alternative predation combined with predator control may be reducing predation pressure on non-game prey where gamebirds are released, but high fox density elsewhere likely results in higher predation pressure on a wide range of species.